基于ZrO_(2)/AuNPs膜修饰电极的适配体传感器用于凝血酶的检测研究  

Sensitive Electrochemical Aptasensor for the Detection of Thrombin based on Zirconia/Nanogold Film Modified Electrode

作　　者：文艳清[1] 龙倩[1] 张友玉[2] 李海涛[2] WEN Yanqing;LONG Qian;ZHANG Youyu;LI Haitao(Safety Technology,Changsha 410151;College of Chemistry and Chemical Engineering,Hunan Normal University,Changsha 410081)

机构地区：[1]湖南安全技术职业学院,湖南长沙410151 [2]湖南师范大学化学化工学院,湖南长沙410181

出　　处：《分析科学学报》2023年第3期331-336,共6页Journal of Analytical Science

基　　金：国家自然科学基金(No.21275051,21475043);湖南师范大学化学生物学及中药分析教育部重点实验室开放基金课题(KLCBTCMR2015-04,KLCBTCMR18-09)。

摘　　要：提出了一种简单、无标记、可再生的电化学方法研究适配体和凝血酶之间的相互作用,采用亚甲基蓝(MB)做电化学指示剂,氧化锆(ZrO_(2))-金纳米粒子(AuNPs)涂层修饰玻碳电极(GCE)。利用金-硫键及杂交化学反应,捕获探针和适配体依次修饰到电极表面,亚甲基蓝插入到DNA上,形成适配体传感器。电极表面的DNA双链在凝血酶的存在下发生解旋,MB在DNA上的吸附量随之减少,峰电流也显著降低,达到检测凝血酶的目的。实验显示,凝血酶在20 pmol/L~150 nmol/L的浓度范围内,峰电流的减小量随凝血酶浓度的升高而增大,检出限为20.6 fmol/L。该方法简单、灵敏、选择性好,并成功用于实际样品检测。In this paper,a simple,label-free and regenerative electrochemical method was proposed to study the interaction between aptamer and thrombin by using methylene blue(MB)as an electrochemical indicator based on zirconia(ZrO_(2))and gold nanoparticles(NG)film modified glassy carbon electrode(GCE).A thiolated capture probe was firstly attached onto the NG/ZrO_(2)/GCE by gold-sulfur affinity.Aptamer probe which was used to partly hybridizie with capture DNA sequence and specifically recognize thrombin,was then immobilized on the electrode surface by hybridization reaction.The interaction of thrombin with the aptamer displaces the aptamer sequence and causes it to dissociate from the interface.This results in a decrease in the amount of aptamer/capture probe duplex form.Accordingly,methylene blue,an electroactive indicator bound to the duplex,is desorped from the electrode surface,which induces the peak current of MB decreased obviously.The peak current of MB linearly decreases with the concentration of thrombin over a range of 20 pmol/L to 150 nmol/L with a detection limit of 20.6 fmol/L.Thus,the fabricated sensor is shown to exhibit high sensitivity.

关 键 词：氧化锆 金纳米粒子 适配体 凝血酶 亚甲基蓝 电化学适配体传感器 

分 类 号：O657.1[理学—分析化学]